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. 2014 Jul 8;9(7):e101969. doi: 10.1371/journal.pone.0101969

Figure 2. Pyrvinium inhibits WNT signaling in CRC cells.

Figure 2

A. Pyrvinium inhibits WNT reporter gene activity in CRC cells. Cells were treated with Pyrvinium at the indicated concentrations and relative WNT reporter luciferase activity determined. Immunoblots show that Pyrvinium decreases PYGOPUS2 levels in both SW480 and HCT116 WTKO cells in a dose-dependent manner. Error bars indicate ± S.E.M of three experiments. B. Expression of WNT regulated genes AXIN2, DKK1, CCND1 and LGR5 in CRC cell lines treated with 100 nM Pyrvinium or vehicle control was determined using real-time RT-PCR. Error bars indicate ± S.E.M of three experiments. *p<0.05. C. A nuclear or cytosolic fraction was extracted from vehicle or 100 nM Pyrvinium treated SW480 and HT29 cells and CTNNB1 protein levels were determined by immunoblot. D. Immunoblot of the WNT associated biomarkers CTNNB1 and CCND1 in CRC cell lines treated with Pyrvinium or vehicle control. Cells were treated, lysed in protein sample buffer and immunoblotted with the cognate antibodies for CTNNB1, CCND1 and GAPDH control.